Premium
Quantitative determination of saquinavir from Caco‐2 cell monolayers by HPLC‐UV
Author(s) -
Ucpinar Sibel Demirbas,
Stavchansky Salomon
Publication year - 2003
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.205
Subject(s) - chemistry , saquinavir , chromatography , high performance liquid chromatography , monolayer , biochemistry , medicine , family medicine , human immunodeficiency virus (hiv) , viral load , antiretroviral therapy
Abstract The validation and quantitative determination of the protease inhibitor, saquinavir, from confluent Caco‐2 monolayers and from aqueous solution is reported. The high performance liquid chromatographic method consisted of an Ultramex™ 5 C 8 reverse‐phase column (250 × 4.6 mm i.d.) and a mobile phase of acetonitrile:water:triethylamine (55:44:1, v/v/v, pH 6.5). Samples were analyzed using an ultraviolet detector at 238 nm, and diltiazem hydrochloride (66 µg/mL) was used as an internal standard. A linear response over a broad concentration range (0.4–8.0 µg/mL, r 2 = 0.997) was obtained. The limit of detection and quantitation was set at 0.14 and 0.4 µg/mL, respectively. Over a 4 day period, the intra‐day and inter‐day precision ranged from 1 to 7% with a mean of 4%, and from 1 to 2% with a mean of 1.5%, respectively. Bench–top and storage stability of saquinavir was found to be satisfactory. The permeability of saquinavir through Caco‐2 monolayers was estimated using this assay. Copyright © 2002 John Wiley & Sons, Ltd.